Coomassie Blue Staining SDS-PAGE Gels

1. Fix gel in 10 volumes of 1X fixer for 30 minutes at room termperature. Fixer (1X) is 40% methanol and 10% acetic acid. Discard fixer when done. This step may go overnight. 2. Stain gel in 10 volumes of Coommassie Blue Stain for 1 hour at room temperature. Save staining solution. Re-use solution for 1 month to 1 year. This step may go overnight. 3. Destain gel in 10 volumes of 1X destain for 30 minutes at room temperature. 4. Repeat the destain step. 5. Soak gel in 1-2% glycerol in ddH2O for 15 minutes. The gel is ready to photograph or dry.

Rehydration of a dried gel: 1. Place dried gel with paper in 100-200 mls of: 30% glycerol 20% methanol 10% acetic acid 2. Rotate or rock gently for 30 minutes to 2 hours at room termperature. 3. After gel has hydrated, it may be removed from paper and equlibrated with solutions with higher water content.

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